Generation of specific inhibitors of SUMO-1– and SUMO-2/3–mediated protein-protein interactions using Affimer (Adhiron) technology

DOI: 10.1126/scisignal.aaj2005

Authors: David J. Hughes (University of Leeds; University of St. Andrews) , Christian Tiede (University of Leeds) , Natalie Penswick (University of Leeds) , Anna Ah-san Tang (University of Leeds) , Chi H. Trinh (University of Leeds) , Upasana Mandal (University of Leeds) , Katarzyna Z. Zajac (University of Leeds) , Thembaninkosi Gaule (University of Leeds) , Gareth Howell (University of Leeds) , Thomas A. Edwards (University of Leeds) , Jianxin Duan (Schrödinger GmbH) , Eric Feyfant (Schrödinger Inc) , Michael J. Mcpherson (University of Leeds) , Darren C. Tomlinson (University of Leeds) , Adrian Whitehouse (University of Leeds)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Science Signaling , VOL 10

State: Published (Approved)
Published: November 2017

Abstract: Because protein-protein interactions underpin most biological processes, developing tools that target them to understand their function or to inform the development of therapeutics is an important task. SUMOylation is the posttranslational covalent attachment of proteins in the SUMO family (SUMO-1, SUMO-2, or SUMO-3), and it regulates numerous cellular pathways. SUMOylated proteins are recognized by proteins with SUMO-interaction motifs (SIMs) that facilitate noncovalent interactions with SUMO. We describe the use of the Affimer system of peptide display for the rapid isolation of synthetic binding proteins that inhibit SUMO-dependent protein-protein interactions mediated by SIMs both in vitro and in cells. Crucially, these synthetic proteins did not prevent SUMO conjugation either in vitro or in cell-based systems, enabling the specific analysis of SUMO-mediated protein-protein interactions. Furthermore, through structural analysis and molecular modeling, we explored the molecular mechanisms that may underlie their specificity in interfering with either SUMO-1–mediated interactions or interactions mediated by either SUMO-2 or SUMO-3. Not only will these reagents enable investigation of the biological roles of SUMOylation, but the Affimer technology used to generate these synthetic binding proteins could also be exploited to design or validate reagents or therapeutics that target other protein-protein interactions.

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography

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